肾癌相关基因GYLZ-RCC18对肾癌细胞系GRC-1抗死亡作用机制的研究

Anti-dead effect of renal cell carcinoma related novel gene GYLZ-RCC18 on renal cell carcinoma cell line GRC-1

目的 应用反义基因转染技术探讨肾癌相关新基因GYLZ RCC18与肾癌细胞死亡 (包括坏死及凋亡 )之间的关系 ,并探讨肾癌基因疗法的新途径。 方法 用脂质体包裹的新基因GYLZ RCC18第一编码区起始处的 2 0个碱基的反义和正义寡核苷酸导入肾癌细胞系GRC 1,应用流式细胞技术和伊红排斥实验连续检测GYLZ RCC18反义寡核苷酸对肾癌细胞凋亡、坏死的影响。 结果 导入GYLZ RCC18反义寡核苷酸后 ,可大量杀伤GRC 1细胞并明显促进GRC 1细胞坏死 ,于第 4天达到杀伤高峰约 6 0 % ;同时可连续 8d诱导凋亡峰 ,最高于第 2天达到约 2 2 5 % ,两种作用曲线形状和峰值不完全一致。 结论 GYLZ RCC18是肾癌相关的重要的新的癌基因 ,GYLZ RCC18的表达可通过两种不同的机制即抗癌细胞坏死机制和抗凋亡机制发挥对肾癌细胞的保护作用 ,GYLZ RCC18可能对肾癌细胞生存和耐药至关重要。通过导入反义基因的方法阻断肾癌细胞中GYLZ RCC18的表达 ,可有效地杀伤肾癌细胞。对此新基因的研究将有助于为肾癌的基础研究和临床治疗提供新的思路和途径。

Objective To transfect the anti se nse oligonuclotide of GYLZ RCC18 into renal cell carcinoma cell line GRC 1 and study its role in the development of RCC. Methods[WT5”B Z] 20 bp GYLZ RCC18 antisense oligonuclotide packed with liposome was transf ected into GRC 1 cell and the changes in apoptosis and mortality of GRC 1 were observed by using flow cytometry and staining. Results[WT5 ”BZ] After transfection of GYLZ RCC18 antisense oligonuclotide, the mortal ity of GRC 1 increased evidently. The antisense oligonuclotide induced the apop tosis of GRC 1. Clonclusions GYLZ RCC18 is o ne of RCC closely related novel gene. It may exert an antidead and antiapoptosi s effect on RCC. It is also related to the multidrug resistance of RCC. Transfec tion of antisense oligonuclotide could inhibite the development of RCC. The stud y of this gene provides a new clue for the study and gene therapy of RCC.